Enhanced binding and killing of target tumor cells by drug-loaded liposomes modified with tumor-specific phage fusion coat protein.
Enhanced binding and killing of target tumor cells by drug-loaded liposomes modified with tumor-specific phage fusion coat protein.
复制标题
DOI：
10.2217/nnm.10.30
复制
发表时间：
2010-06
期刊：
影响因子：
--
通讯作者：
Torchilin VP
中科院分区：
文献类型：
--
作者：
Wang T;D'Souza GG;Bedi D;Fagbohun OA;Potturi LP;Papahadjopoulos-Sternberg B;Petrenko VA;Torchilin VP
目的：探索利用噬菌体展示技术鉴定的癌细胞特异性噬菌体融合pVIII外壳蛋白，用于靶向向MCF-7乳腺癌细胞输送载药脂质体。利用8聚体景观文库f8/8和针对MCF-7细胞的生物扫描方法筛选出含有MCF-7特异性多肽的景观噬菌体蛋白。用动态光散射法和冷冻切片电子显微镜对肿瘤特异性噬菌体融合外壳蛋白修饰的阿霉素脂质体的大小和形态进行了研究。用免疫印迹法检测脂质体中噬菌体蛋白的拓扑结构。用荧光显微镜和荧光分光光度法检测噬菌体多西与MCF-7细胞的结合作用。利用靶细胞和非靶细胞共培养模型，流式细胞仪显示了对MCF-7的选择性靶向。CellTiter-Blue®检测和caspase-3/CPP32荧光检测证实噬菌体-Doxil诱导的肿瘤细胞杀伤和凋亡。从噬菌体景观库中鉴定出针对MCF-7细胞的嵌合噬菌体融合外壳蛋白，将其直接掺入载阿霉素的聚乙二醇化脂质体(Doxil®)的脂质体双层中，而不需要与亲脂部分进行额外的连接。Western blotting证实在噬菌体Doxil中同时存在靶向肽和pVIII外壳蛋白，保持了脂质体的形态，并在噬菌体蛋白掺入后保留了大部分被掺入的药物。噬菌体融合pVIII外壳蛋白与脂质体结合后仍保持结合活性，噬菌体-多西环素对MCF-7细胞具有较强的特异性靶向性，体外对靶细胞的杀伤作用显著增强。我们提出了一种新的和直接的方法，通过在其表面锚定特定的噬菌体蛋白(替代抗体)来制造肿瘤靶向纳米药物。
To explore cancer cell-specific phage fusion pVIII coat protein, identified using phage display, for targeted delivery of drug-loaded liposomes to MCF-7 breast cancer cells. An 8-mer landscape library f8/8 and a biopanning protocol against MCF-7 cells were used to select a landscape phage protein bearing MCF-7-specific peptide. Size and morphology of doxorubicin-loaded liposomes modified with the tumor-specific phage fusion coat protein (phage–Doxil) were determined by dynamic light scattering and freeze-fraction electron microscopy. Topology of the phage protein in liposomes was examined by western blot. Association of phage–Doxil with MCF-7 cells was evaluated by fluorescence microscopy and fluorescence spectrometry. Selective targeting to MCF-7 was shown by FACS using a coculture model with target and nontarget cells. Phage–Doxil-induced tumor cell killing and apoptosis were confirmed by CellTiter-Blue® Assay and caspase-3/CPP32 fluorometric assay. A chimeric phage fusion coat protein specific towards MCF-7 cells, identified from a phage landscape library, was directly incorporated into the liposomal bilayer of doxorubicin-loaded PEGylated liposomes (Doxil®) without additional conjugation with lipophilic moieties. Western blotting confirmed the presence of both targeting peptide and pVIII coat protein in the phage–Doxil, which maintained the liposomal morphology and retained a substantial part of the incorporated drug after phage protein incorporation. The binding activity of the phage fusion pVIII coat protein was retained after incorporation into liposomes, and phage–Doxil strongly and specifically targeted MCF-7 cells, demonstrating significantly increased cytotoxicity towards target cells in vitro. We present a novel and straightforward method for making tumor-targeted nanomedicines by anchoring specific phage proteins (substitute antibodies) on their surface.
登录
查看更多内容
影响因子：
16.1
作者：
Sergeeva, Anna;Kolonin, Mikhail G.;Molldrem, Jeffrey J.;Pasqualini, Renata;Arap, Wadih
通讯作者：
Arap, Wadih
影响因子：
2.9
作者：
PATERNOSTRE, MT;ROUX, M;RIGAUD, JL
通讯作者：
RIGAUD, JL
DOI：
10.1111/j.1432-1033.1997.t01-1-00168.x
发表时间：
1997-11-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Angrand, M;Briolay, A;Roux, B
通讯作者：
Roux, B
影响因子：
3.4
作者：
Stopar, David;Spruijt, Ruud B.;Hemminga, Marcus A.
通讯作者：
Hemminga, Marcus A.
影响因子：
11.2
作者：
Krag, David N.;Shukla, Girja S.;Thomas, Christian
通讯作者：
Thomas, Christian